WO2012067224A1 - Therapeutic or prophylactic agent for corneal epithelium disorders and/or conjunctival epithelium disorders - Google Patents

Abstract

The present invention provides a prophylactic or therapeutic medicinal product or a supplement which is effective on corneal epithelium disorders and/or conjunctival epithelium disorders and is safe. Specifically provided is a therapeutic or prophylactic agent for corneal epithelium disorders and/or conjunctival epithelium disorders, which is characterized by comprising, as an active ingredient, eicosapentaenoic acid and/or docosahexaenoic acid, or a glycerin ester or a phospholipid which contains the fatty acid as a constituent fatty acid, or a lower-alcohol ester of the fatty acid. Preferably, the therapeutic or prophylactic agent contains EPA and/or DHA in the form of a purified fish oil or a purified krill oil. When the therapeutic or prophylactic agent is ingested at a daily dose of about 50-5000 mg in terms of the total amount of EPA and/or DHA or the ester thereof, corneal epithelium disorders and/or conjunctival epithelium disorders can be alleviated or the amount of tear can be recovered.

Description

Treatment or prevention agent for corneal epithelial disorder and / or conjunctival epithelial disorder

The present invention relates to treatment / prevention of eye diseases such as corneal epithelial disorder and / or conjunctival epithelial disorder caused by dry eye, contact lens, irritation by eyelid club lashes, conjunctival stone, viral conjunctivitis, and allergic conjunctivitis. About.

With the spread of personal computers and mobile phones, an increasing number of people are apt to get tired or feel somehow uncomfortable. In particular, in contact lens users, there are an increasing number of cases complaining of discomfort such as dry eyes and groaning by staring at the screen. The cause of such symptoms is dry eyes and dry eyes.
Dry eye is a disease that causes damage to the surface of the eye due to decreased tears or qualitative changes. Lack of tears makes the eyes dry and vulnerable, making contact lenses difficult to use.
According to the dry eye diagnostic criteria by the 2006 Dry Eye Study Group, subjective symptoms (assessment of pain and dryness by VAS test), abnormal tears (evaluated by tear film destruction time or Schirmer test), disorder of keratoconjunctiva When three (evaluation by fluorescein or rose bengal staining) are confirmed, dry eye is diagnosed, and in two cases, it is diagnosed as suspicious.

As symptomatic treatment for dry eye, viscous eye drops such as saline eye drops and chondroitin sulfate are used. Another treatment is to close the tear outlet to keep the tears on the surface of the eye for as long as possible.
In addition to dry eye, corneal epithelium and / or conjunctival epithelium can be finely damaged due to contact lenses, irritation by eyelid varus eyelashes, conjunctival stones, viral conjunctivitis, and allergic conjunctivitis.

Patent Document 1 describes a dry eye nutritional supplement using GLA-rich linseed oil in combination with eicosapentaenoic acid (hereinafter referred to as EPA) or docosahexaenoic acid (hereinafter referred to as DHA). Non-Patent Document 1 describes the effect of essential fatty acids on dry eye.

Special table 2005-535733

Dry eyes, even if not severe, impair the comfort of everyday life. It is an object of the present invention to investigate the cause of dry eye and to provide a safe treatment method that is not symptomatic and does not cause side effects.

The present inventors have found that the amount of tears is reduced in n-3 fatty acid-deficient mice while conducting research on mice lacking n-3 fatty acids, and n-3 fatty acids are involved in tear volume. I thought. Furthermore, it was found that the decrease in tear volume can be returned to the normal level by administering purified fish oil containing a lot of n-3 fatty acids EPA and DHA to mice lacking n-3 fatty acids. Completed the invention.

The gist of the present invention is the therapeutic and / or preventive agent for corneal epithelial disorder and / or conjunctival epithelial disorder of (1) to (7), and the tear recovery agent of (8) to (13).
(1) A therapeutic and / or prophylactic agent for corneal epithelial disorder and / or conjunctival epithelial disorder, characterized by containing EPA and / or DHA or an ester thereof as an active ingredient.
(2) Corneal epithelial disorder and / or conjunctival epithelial disorder is a disorder caused by any of dry eye, contact lens, irritation by eyelid varus eyelashes, conjunctival stone, viral conjunctivitis, and allergic conjunctivitis ( The therapeutic and / or prophylactic agent of 1).
(3) The therapeutic and / or preventive agent according to (1) or (2), wherein the ester of EPA and / or DHA is contained as a glycerin ester, a lower alcohol ester, a phosphate ester or a purified fish oil.
(4) The treatment and / or prevention according to any one of (1) to (3), wherein the total amount of eicosapentaenoic acid and / or docosahexaenoic acid, or an ester thereof is ingested at least about 50 to 5000 mg as a daily dose. Agent.
(5) The therapeutic and / or prophylactic agent according to any one of (1) to (3), in which EPA and / or DHA, or a total amount of these esters is ingested at least about 300 to 3000 mg as a daily dose.
(6) The therapeutic and / or prophylactic agent according to any one of (1) to (3) for ingesting at least about 1000 to 2000 mg of EPA and / or DHA or a total of these esters as a daily dose.
(7) The therapeutic and / or prophylactic agent according to any one of (1) to (6), further comprising tocopherol as an antioxidant.

(8) A tear recovery agent comprising EPA and / or DHA or an ester thereof as an active ingredient.
(9) The tear amount restoring agent according to (7), wherein the ester of EPA and / or DHA is contained as glycerin ester, lower alcohol ester, phosphate ester or purified fish oil.
(10) The tear recovery agent according to (8) or (9), wherein a daily dose of eicosapentaenoic acid and / or docosahexaenoic acid, or a total amount of these esters is taken at least about 50 to 5000 mg.
(11) The tear amount restoring agent according to (8) or (9), wherein the daily amount of EPA and / or DHA, or a total amount of these esters is ingested at least about 300 to 3000 mg.
(12) The tear amount restoring agent according to (8) or (9), wherein the daily amount of EPA and / or DHA, or a total amount of these esters is ingested at least about 900 to 2100 mg.
(13) The tear recovery agent according to any one of (8) to (12), further comprising tocopherol as an antioxidant.

By administering n-3 fatty acids EPA and DHA, corneal epithelial disorder and / or by dry eye, contact lens, irritation by eyelid club lash, conjunctival stone, viral conjunctivitis, and allergic conjunctivitis Conjunctival epithelial disorder can be treated or prevented. Since EPA and DHA can increase tear volume, these symptoms can be alleviated. It is possible to provide pharmaceuticals, supplements, etc. that can reduce symptoms such as dry eye made of refined fish oil that has abundant dietary experience and is free of side effects.

FIG. 1 is a diagram showing the result of evaluating the subjective evaluation of eye pain by the VAS test. In the figure, * P <0.05 and #P <0.1 indicate significant differences according to Mann-Whitney U test. FIG. 2 is a diagram showing the result of evaluating the subjective evaluation of dry eyes by the VAS test. FIG. 3 is a diagram showing the results of measuring Tear film breakup time (tear film breaking time). n = 24 or 30. In the figure, * P <0.05 and #P <0.1 indicate significant differences according to Mann-Whitney U test. FIG. 4 is a diagram showing the results of evaluating eye wounds with fluorescein staining. n = 24 or 30. FIG. 5 is a diagram showing the amount of change in FIG. FIG. 6 is a diagram showing the results of evaluation of eye scratches with rose bengal staining. n = 24 or 30. FIG. 7 is a diagram showing the amount of change in FIG. In the figure, * P <0.05 indicates a significant difference by Mann-Whitney U test. In the figure, ** P <0.05 indicates a significant difference due to two-way ANOVA. FIG. 8 is a diagram showing the results of evaluating tear volume by the Schirmer test. n = 24 or 30.

The present invention is a preventive or therapeutic agent for corneal epithelial disorder and / or conjunctival epithelial disorder comprising EPA and / or DHA or an ester thereof as an active ingredient. There are some reports of using n-6 fatty acids such as linseed oil for the treatment of dry eye. What is the intake of n-3 fatty acids (especially EPA, DHA) alone for corneal epithelial disorder and / or conjunctival epithelial disorder? It is not known whether it has an impact.
The present inventors have found that deficiency of n-3 fatty acids clearly leads to a decrease in tear volume, and have confirmed that such tear volume can be recovered by taking purified fish oil containing EPA and DHA.

The EPA and / or DHA used in the present invention is preferably an ester with glycerin, i.e. triglyceride, diglyceride, monoglyceride or phospholipid, i.e. phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol, cardiolipin, Examples thereof include phosphatidic acid (both including lyso form), or an ester with a lower alcohol (having 1 to 5 carbon atoms), that is, methyl ester, ethyl ester and the like. Preference is given to those containing n-3 fatty acids as the main component, such as EPA and DHA, and substantially free of n-6 fatty acids. Specifically, the total of n-6 fatty acids is 10% by weight or less, preferably 5% by weight or less.
Specifically, as the triglyceride, refined fish oil, concentrated oil (enriched EPA and DHA concentration of refined fish oil by enzyme reaction or chemical reaction) and the like can be used. Moreover, krill oil etc. can be used as a phospholipid. Examples of n-3 polyunsaturated fatty acid sources containing EPA and DHA include seafood extracts, animal extracts, egg yolk extracts, plant extracts, fungal extracts, etc., preferably krill oil, fish oil, fish extracts, Examples include squid extract, skipjack ovary extract, genetically modified plant extract, and the like. Examples of materials containing a large amount of phospholipid include krill oil, squid extract and bonito ovary extract. The phospholipid concentration and purity in these materials can be arbitrarily adjusted by using techniques generally known in the art such as concentration, extraction, and / or purification, and blending.
EPA and / or DHA are known to be effective for hyperlipidemia and allergic symptoms, and many pharmaceuticals and various supplements are already on the market. For the purposes of the present invention, capsule preparations similar to these can be used. It can also be added to various beverages and foods.
Specifically, a product obtained by processing a fat and oil obtained by adding an antioxidant to purified fish oil into a gelatin capsule is often used.

As the refined fish oil, any fish oil may be used as long as it contains EPA and DHA. Examples of fish oils rich in EPA include sardine oil and cod liver oil, and examples of fish oils rich in DHA include tuna oil and bonito oil.
EPA / DHA concentrated oil is oil and fat with increased EPA / DHA concentration in purified fish oil triglycerides using regioselective, fatty acid-selective lipases, etc. (see Japanese Patent Publication No. 4-16519 etc.) ).

The therapeutic and / or prophylactic agent of the present invention comprises EPA and / or DHA or an ester thereof per day as a total amount of EPA and DHA of about 50 to 5000 mg, preferably 300 to 3000 mg, more preferably about 900 to 2100 mg, particularly preferably 1100-1900 mg, is orally administered once or divided to a patient with dry eye symptoms. The effect is observed by continuous administration for at least 4 weeks.
EPA and / or DHA already have long experience in other applications, so there are no safety issues.

In the corneal epithelial disorder and / or conjunctival epithelial disorder treatment and / or prevention agent or tear recovery agent of the present invention, anthocyanidin, lutein, eyebright, megsurinoki, vitamins and the like, which are preferable for eye fatigue recovery, are added. And may be used in combination.

Examples of the present invention will be described below, but the present invention is not limited to these.

Production of n-3 fatty acid-deficient animals Generally, laboratory animal feeds contain n-3 fatty acids containing an appropriate amount of DHA. It is necessary to use a special feed that does not contain fatty acids. In addition, normally reared animals ingest sufficient DHA from the placenta and mother's milk during the period from prenatal to weaning, and it is not easy to produce the target n-3 fatty acid deficient animals in the first generation. Absent. Taking these into account, n-3 fatty acid deficiency containing 7% lipids based on AIN93G (standard purified feed composition for nutritional research for mice and rats published by the National Institute of Nutrition) A feed (n-3 Def, linoleic acid, 14.4%; α-linolenic acid, 0.1%) was prepared. As a normal feed, an n-3 fatty acid-containing feed containing α-linolenic acid (18: 3n-3) (n-3 Adq, linoleic acid, 14.3%; α-linolenic acid, 2.6%) was used. These feeds were fed to female mice immediately after weaning (3 weeks of age), bred, mated and then mated to obtain the second generation, and this second generation of 11-month-old male mice were used in the experiment . Moreover, in order to avoid the influence of the mother animal in the experiment, all the animals of each group were set so that the mother animal was different.

Preparation of test drug The test drug is fish oil (manufactured by Nippon Suisan Co., Ltd., refined fish oil EPA-28 (EPA 28% or higher, DHA 12% or higher)). DHA amount is 5 mg / head and EPA amount is 11 Prepared to be mg / head.
As a control group, coconut oil was used, and it was set to be equal to the amount of fatty acid of the fish oil group. The main fatty acid composition of the fish oil used is shown in Table 1 (the content of 18: 2 which is an n-6 fatty acid was 1.1%, and 20: 4 was 1.3%).

Operating procedure Using tear-liquid phenol red thread (Zone-Quick, Showa Yakuhin), the amount of tears secreted in 30 seconds by n-3 Def and n-3 Adq mice (the length of red thread) It was measured. Divide these animals into two groups, and orally administer fish oil or coconut oil-containing diet for 7 days, measure tear volume again on the 8th day, retina related to vision on the 9th day, lacrimal gland related to lacrimal secretion Meibomian glands were collected and the fatty acid composition was measured.

Experimental Results and Discussion The tear fluid is composed of three layers, the oil layer secreted from the meibomian gland, the water layer secreted from the lacrimal gland, and the mucin layer secreted from the conjunctiva in order from the one in contact with the outside.
Dry eye, also known as dry cornea, is a type of eye disease that causes eye discomfort and abnormal visual function due to dryness of the surface of the eyeball, and tears that reduce the lacrimal fluid secreted from the lacrimal gland There are known a decrease in liquid type and an increased tear evaporation type that occurs because the oil layer secreted from the meibomian glands is not sufficient.
The amount of tear fluid in n-3 fatty acid-deficient aged mice showed a clear decrease, and was judged to exhibit dry eye symptoms (Table 2). In the fish oil group in which fish oil was administered to these n-3 fatty acid-deficient mice for 7 days, a significant increase in tear volume (recovery of tear volume) was observed compared to the control group (Table 3). Moreover, since the tear volume was comparable to that of normal mice, the tear volume seemed to have almost completely recovered. In the EPA and DHA fatty acid composition results of the organs involved in tear volume, an increase was observed in both the lacrimal gland and meibomian gland by administration of fish oil (Table 4).
From the above results, it became clear that n-3 fatty acid deficient animals showed clear dry eye symptoms and that the symptoms were improved by ingestion of fish oil. In addition, it was considered that the dry eye symptom of n-3 fatty acid-deficient animals was a decrease in lacrimal secretion due to decreased functionality of either the lacrimal gland, the meibomian gland, or either, based on the reaction results of fatty acid composition after fish oil administration.

 

Using the same fish oil as in Example 1 (manufactured by Nippon Suisan Co., Ltd., refined fish oil EPA-28 (EPA 28% or more, containing DHA 12% or more)), the effect on human corneal epithelial disorder was confirmed.

Study Protocol Summary The efficacy of EPA and DHA was evaluated in a double-blind study on 27 subjects diagnosed with dry eye according to the dry eye diagnostic criteria (2006 Dry Eye Study Group). Active or Placebo supplements were ingested for 12 weeks with normal dry eye treatment such as eye drops. The active group was administered purified fish oil capsules (15 capsules) in an amount to consume 1245 mg of EPA and 540 mg of DHA per day. In the Placebo group, medium-chain fatty acid triglyceride (MCT) capsules (15 capsules) were administered. The efficacy was measured 5 times before administration, 4 weeks after administration, 8 weeks, 12 weeks, and 4 weeks after administration.

The following 6 items were evaluated.
・ VAS test subjective evaluation of eye pain: Visual analog
Use scale to visually evaluate the degree of pain.
・ VAS test subjective evaluation of dry eyes: Visual analog
Use scale to visually assess the degree of dryness.
・ Tear film breakup time: One of the diagnostic criteria for dry eye. A test that measures the time it takes for the tear film on the corneal surface to break down after the instillation of fluorescein. Determine tear retention. Assess normal for 10 seconds or more and suspect dry eye for 5 seconds or less.
・ Evaluate the wound of the eyes with fluorescein staining: A yellow stain called fluorescein is instilled to stain the wound on the surface of the eye, and the severity of dry eye is determined. Fluorescein also dyes the conjunctiva, but can primarily assess corneal wounds.
・ Evaluate wounds of eyes with rose bengal staining: A red staining solution called rose bengal is instilled to stain the wounds on the surface of the eye to determine the severity of dry eye. Rose Bengal can mainly evaluate conjunctiva in which the mucin layer is impaired.
Evaluate tear volume with the Schirmer test: The Schirmer test is a test for measuring the amount of tears secreted, and a long filter paper is sandwiched between the lower eyelids for 5 minutes and the length wet with tears is measured. Evaluate 5mm or less as an abnormal value.

Results The number of subjects was 12 in the Active group and 15 in the Placebo group.
The results of each evaluation item are shown in FIGS.
The evaluation of eye pain by the VAS test, tear film destruction time, and the Rose Bengal staining test showed that there were statistically significant effects compared to the Placebo group. In other items, the Active group showed a tendency to be better than the Placebo group.

Capsules containing active group and medium-chain fatty acid triglycerides, in which EPA 162 mg, DHA 784 mg, anthocyanidin 59 mg and lutein 17 mg are ingested as capsules per day in a double-blind randomized controlled trial for subjects with eyestrain symptoms The effect was compared with the Placebo group that received the drug.

Results The number of subjects was 11 in the Active group and 9 in the Placebo group. In the subjective symptom evaluation 4 weeks after the start of administration, among the ocular symptoms, a statistically significant recovery was observed in the Active group compared to the Placebo group in the dry item. Among the symptoms of the eyes, in the items of fatigue, haze, hyperemia, and flicker, the Active group showed a significant recovery 4 weeks after the start of administration compared to before administration.

According to the present invention, dry eye, contact lens use, irritation caused by eyelid varus eyelashes, conjunctival stones, viral conjunctivitis, allergic conjunctivitis and other causes of eyes, especially corneal epithelium and conjunctival epithelium are alleviated, discomfort etc. We can supply medicines and supplements to reduce the risk.

Claims (13)

1. Corneal epithelial disorder and / or conjunctival epithelium characterized by containing, as an active ingredient, eicosapentaenoic acid and / or docosahexaenoic acid, glycerin ester or phospholipid containing these fatty acids as constituent fatty acids, or lower alcohol esters of these fatty acids A therapeutic and / or prophylactic agent for disorders.
2. The corneal epithelial disorder and / or the conjunctival epithelial disorder is a disorder caused by any one of dry eye, contact lens, irritation by eyelid varus eyelashes, conjunctival stone, viral conjunctivitis, and allergic conjunctivitis. Treatment and / or prevention agent.
3. The therapeutic and / or preventive agent according to claim 1 or 2, comprising glycerin ester or phospholipid containing eicosapentaenoic acid and / or docosahexaenoic acid as a constituent fatty acid as purified fish oil or purified krill oil.
4. The therapeutic and / or prophylactic agent according to any one of claims 1 to 3, for taking about 50 to 5000 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
5. The therapeutic and / or prophylactic agent according to any one of claims 1 to 3, for taking about 300 to 3000 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
6. The therapeutic and / or prophylactic agent according to any one of claims 1 to 3, for taking about 900 to 2100 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
7. The therapeutic and / or prophylactic agent according to any one of claims 1 to 6, further comprising tocopherol as an antioxidant.
8. (2) A tear recovery agent comprising eicosapentaenoic acid and / or docosahexaenoic acid, or a glycerin ester or phospholipid containing these fatty acids as constituent fatty acids, or a lower alcohol ester of these fatty acids as an active ingredient.
9. The tear recovery agent according to claim 8, comprising glycerin ester or phospholipid containing eicosapentaenoic acid and / or docosahexaenoic acid as a constituent fatty acid as purified fish oil or purified krill oil.
10. 10. The tear recovery agent according to claim 8 or 9, for taking about 50 to 5000 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
11. 10. The tear volume recovery agent according to claim 8 or 9, for taking about 300 to 3000 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
12. 10. The tear volume recovery agent according to claim 8 or 9, for taking about 900 to 2100 mg of eicosapentaenoic acid and / or docosahexaenoic acid as a daily dose.
13. The tear recovery agent according to any one of claims 8 to 12, further comprising tocopherol as an antioxidant.
    

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